Quantitative PCR (qPCR) is widely used to detect viruses. However, mismatches occurring in the 3'-end of the primers reduce amplification efficiency of qPCR and limit its capacity in detection of highly variable viruses. Here, we reported a mismatch-tolerant RT-qPCR with a small amount of additional high-fidelity DNA polymerase for simultaneous detection of RSV-A and RSV-B. The novel assay had higher amplification efficiency for various variants forming mismatches with the primers than the conventional RT-qPCR, and showed good specificity and sensitivity. It demonstrated a good correlation coefficient with a commercial RSV detection kit and had relatively lower Ct values than the kit for 16 of 20 RSV-positive samples. The mismatch-tolerant qPCR technique is a promising approach for sensitive detection of highly variable viruses.

中文翻译：

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耐错配的RT定量PCR：在呼吸道合胞病毒的广谱检测中的应用。

定量PCR（qPCR）被广泛用于检测病毒。但是，在引物3'端出现的错配会降低qPCR的扩增效率，并限制其检测高度可变病毒的能力。在这里，我们报道了具有少量其他高保真DNA聚合酶的错配耐受RT-qPCR，可同时检测RSV-A和RSV-B。与传统的RT-qPCR相比，该新方法对与引物形成错配的各种变异具有更高的扩增效率，并显示出良好的特异性和敏感性。与市售RSV检测试剂盒相比，它显示出良好的相关系数，并且相对于20种RSV阳性样品中的16种，试剂盒的Ct值相对较低。耐错配的qPCR技术是灵敏检测高度可变病毒的一种有前途的方法。